DNA-directed peptide synthesis. VI. Regulating the expression of the lac operon in a cell-free system.

The operon model was proposed by Jacob and Monod in 1961 to explain the regulation of enzyme synthesis in bacteria. The model requires two genetic elements: a locus i which directs the synthesis of a repressor, and a locus called the operon which contains an operator gene and associated structural genes. The repressor is believed to stop RNA and enzyme synthesis by combining with the operator site. The inhibiting effect of the repressor is reversed by an inducer which interacts directly with the repressor. A DNA-dependent cell-free system has been developed for studying the regulation of the operon containing the gene for beta-galactosidase enzyme. In this cell-free system, gene activity is indirectly measured by the synthesis of part of the enzyme. Qualitatively, the regulation of the operon in the cell-free system is similar to its regulation in intact cells. The cell-free system is used to advantage in experiments where it is desirable to use predetermined levels of repressor, operon, and inducer. Such quantitative studies suggest that two inducer molecules are most effective in removing the repressor from the gene.